Use of B-lapachone for non-small cell lung cancer (NSCLC) therapy Lung cancer is the leading cause of death by cancer in the U.S. Non-small cell lung cancer (NSCLC) comprises over 75% of lung cancers diagnosed. Current chemotherapeutic regimens are not effective against NSCLC, with five year survival rates hovering at a mere 14%. Regimens that exploit cancer-specific targets, specifically in and increase therapeutic indices, should enhance the survival rates of patients. We hypothesize that B-lapachone (B-lap), a drug that selectively kills cancer cells with elevated levels of the two-electron reductase, NAD(P)H:quinone oxidoreductase 1 (NQOI), will be an effective agent for use against NSCLC cells that specifically over-express this bioactivating enzyme. Cell death caused by B-lap is not dependent on cell cycle status, not dependent on p53, pRb, or caspases, and downstream cell death events are consistent with calpain- mediated apoptosis. More importantly, cell death by B-lap is dependent on NQO1 expression, where NQO1- deficient cells are resistant to the drug, correction of cells with NQO1 restores lethality, and co-administration of dicoumarol (an NQO1 inhibitor) prevents lethality. NQOI is typically elevated 4- to >100-fold in human NSCLC, indicating the use of B-lap for treatment of this disease. Recent development of novel drug delivery methods make it feasible to administer this drug to determine efficacy against NSCLC in animal models. Three aims will test this hypothesis: Aim #1: Evaluate the role of NQO1 in B-lap-mediated cell death in NSCLC cells, and develop lab correlates for use in future therapy. (Years 1-3). Aim #2: Develop drug vehicles for B-lap that either deliver the drug locally to the lung, or utilize systemic delivery schemes that allow accumulation of the vehicle-drug complexes within the lung, while offering simultaneous treatment for metastatic disease. (Years 1-5). Ail!l._: Compare B-lap-encoded microparticles developed in Aim #2 for specific delivery to the lung to systemic delivery (i.p.) of HP-B-cyclodextrin-B-lap complexes for NQOl-specific lung tumor responses. We will combine these therapies with systemic dicoumarol administration for normal tissue protection. (Years 1-5). We have assembled a strong research team with the needed experience to develop novel drug vehicles, image the deposition and delivery of both B-lap and the vehicle in the lung, and test the hypothesis that B-lap should be an efficacious agent against NSCLC due to tumor-specific elevation of NQO 1, a bioactivating enzyme specifically needed for novel calpain-mediated cell death responses elicited by B-lapachone. We will examine the possibility that co-administration of dicoumarol can act as an antidote, increasing the anti-tumor efficacy of B-lap. PERSONNEL ENGAGED ON PROJECT, INCLUDING information in the format shown below on a/l individuals participating David A. Boothman, Ph.D., Jinming Gao, Ph.D. Zhenghong Lee, Ph.D. Steven Dubinett, M.D. William G. Bornmann, Ph.D. Charles Hoppel, M.D. Erik Bey, Ph.D. At Yu, Ph.D. Sharven Sherma, Ph.D. Melissa Bentle Andrew Wiedmann Andrew Bruening CONSULTANTS/COLLABORATORS. Use continuation pages as needed to provide the required in the project. Principal Investigator (PI) Co-Investigator Co-Investigator PI, Subcontract Consultant Consultant Research Associate Research Associate Consultant Graduate Student, Pharmacology Graduate Student, Biomedical Engineering Research Assistant II PHS 398 (Rev. 9/91) Page 2 BB CC PrincipalnvestigatodProgramDirector (Last.First. Middle): Boothman. David. A.. Ph.D. RESEARCH GRANT TABLE OF CONTENTS PAGE NUMBERS Face Page .................................................................................................................................... 1 Description (Abstract) and Personnel .............................................................................................. 2 Table of Contents .................................................................................................................... 3 Detailed Budget for Initial Budget Period ......................................................................................... 4 Budget for Entire Proposed Project Period & Budget Justification ..................................................... 5 Budgets Pertaining to Consortium/Contractual Arrangements .......................................................... 6-7 Biographical Sketch-Principal Investigator/Program Director (Pages 8-13 intentionally left blank).. 14-17 Other Biographical Sketches .......................................................................................................... 18-42 Resources and Environment .......................................................................................................... 43-51 Research Plan (pgs. 22-46) Introduction to Revised Application (Not to exceed three pages) ...................................................... Introduction to Supplemental Application (Not to exceed one page) .................................................. - 1. Hypothesis and Specific Aims ............................................................................................ 52-53 2. Background and Significance (Introduction) ........................................................................ 53-61 3. Progress Report/Preliminary Studies/Significance (Not to exceed 25 pages*) ..................... 61-67 4. Research Design and Specific Methods .............................................................................. 67-76 5. Human Subjects ................................................................................................................ 77 6. Vertebrate Animals ............................................................................................................ 77 7. Consultants/Collaborators .................................................................................................. 77 8. Consortium/Contractual Arrangements ............................................................................... 77a 9. Literature Cited (Not to exceed six pages) ......................................................................... 78-88 Checklist ...................................................................................................................................... 89 *Type density and size must conform to limits provided in Specific Instructions on page 10. Appendix (Five collated sets. No page numbering necessary for Appendix) Number of publications and manuscripts acceptedor submittedfor publication (Not to exceed 10): 5 Other items (list): _heck if Appendix is Included. Item #1: List of Abbreviations Item #2: Relevant Publications 1. Pink, J.J., Planchon, S.M., Tagliarino, C., Varnes, M.E., Siegel, D., & Boothman, D.A.J. Biol. Chem., 275 (8): 5416-22, 2000. 2. Tagliarino, C., Pink, J.J., Dubyak, G.R., Nieminen, A-L., & Boothman, D.A.J. Biol. Chem. 276(22): 19150-19159, 2001. 3. Planchon, S.M., Pink, J.J., Tagliarino,, C., Bornmann, W.G., Varnes, M.E., & Boothman, D.A. Exp. Cell Res., 267: 95-106, 2001. 4. Nasongkla, N., Wiedmann, A.F., Bruening, A., Beman, M., Bornmann, W.G., Boothman, D.A., and Gao, J., Enhancement of solubility and bioavailability offi-lapachone using cyclodextrin inclusion complexes., Submitted, 2002. 5. Lee, Z., and Berridge, M.S. PET imaging-based evaluation of aerosol drugs and their delivery devices: Nasal and pulmonary studies. IEEE Transactions on Medical Imaging, In Press, 2002. PHS 398 Page 3 CC